Aavrh74 particles for gene therapy of muscle disease

Abstract

Provided herein are AAVrh74 capsid proteins comprising one or more amino acid substitutions or deletions that confer a liver-detargeting property to AAV particles comprising them. Provided herein are AAVrh74 capsid proteins comprising one or more amino acid substitutions that confer an improved transduction efficiency (e.g., in muscle cells). Also provided herein are methods of using AAVrh74 capsid proteins comprising one or more amino acid substitutions or deletions.

What is claimed is: 
     
         1 . An AAVrh74 capsid protein comprising an amino acid substitution or deletion at a position corresponding to T265 of wild-type AAVrh74 capsid protein of SEQ ID NO: 1. 
     
     
         2 . The AAVrh74 capsid protein of  claim 1 , further comprising an amino acid substitution at a position corresponding to Y447, T494, K547, N665, and/or Y733 of wild-type AAVrh74 capsid protein of SEQ ID NO: 1. 
     
     
         3 . The AAVrh74 capsid protein of  claim 1 or claim 2 , wherein the amino acid corresponding to T265 of wild-type AAVrh74 capsid protein of SEQ ID NO: 1 is deleted. 
     
     
         4 . The AAVrh74 capsid protein of  claim 1 or claim 2 , wherein the amino acid corresponding to T265 of wild-type AAVrh74 capsid protein of SEQ ID NO: 1 is substituted. 
     
     
         5 . The AAVrh74 capsid protein of  claim 4 , wherein substitution at the amino acid position corresponding to T265 of wild-type AAVrh74 capsid protein of SEQ ID NO: 1 is T265D, T265F, or T265G. 
     
     
         6 . The AAVrh74 capsid protein of  claim 1 , wherein the capsid protein comprises an amino acid substitution or deletion at a position corresponding to T265 and amino acid substitutions at positions corresponding to:
 (a) Y447 and Y733; or   (b) Y447, Y733, and T494   of wild-type AAVrh74 capsid protein of SEQ ID NO: 1.   
     
     
         7 . The AAVrh74 capsid protein of  claim 6 , wherein the capsid protein comprises a deletion or one of the following substitutions at a position corresponding to T265: T265D, T265F, and T265G, and substitutions corresponding to:
 (a) Y447F and Y733F; or   (b) Y447F, Y733F, and T494T   of wild-type AAVrh74 capsid protein of SEQ ID NO: 1.   
     
     
         8 . A nucleic acid encoding the AAVrh74 capsid protein of  any one of the preceding claims . 
     
     
         9 . An AAV particle comprising the AAVrh74 capsid protein of any one of  claims 1-7 , and a nucleic acid comprising a gene of interest. 
     
     
         10 . The AAV particle of  claim 9 , wherein the nucleic acid further comprising a muscle-specific promoter. 
     
     
         11 . The AAV particle of  claim 9 or claim 10 , wherein the gene of interest encodes a therapeutic protein. 
     
     
         12 . The AAV particle of  claim 11 , wherein the therapeutic protein is dystrophin, myotilin, lamin, caveolin, caplain-3, dysferlin, a sarcoglycan, AUF1, TCAP, TRIM32, FKRP, titin, acetylflucosamine epimerase, Desmin, LARGE, fukutin, an integrin, salenoprotein, a collagen, plectin, or a functional fragment thereof. 
     
     
         10 . A composition comprising the AAV particle of any one of claims  9 - 12  and a pharmaceutically acceptable carrier. 
     
     
         11 . A method comprising administering to a subject the composition of  claim 10 . 
     
     
         12 . The method of  claim 11 , wherein the subject is a human subject. 
     
     
         13 . The method of any  claim 11 or 12 , wherein the subject suffers from or is at risk of suffering from a muscular dystrophy.